Biochemistry in situ to determine inheritance of RNA-protein complexes The long-term goal is to elucidate the molecular mechanisms how macromolecular RNA-protein complexes transmit information to future generations of cells and progeny. We classically think of DNA and DNA modifications as the only information inherited between cells. Recent work demonstrates that RNA and RNA- binding proteins are also inherited and that these proteins have functions in organism development and immunity. My lab aims to identify which RNA binding proteins are inherited, determine the macromolecular organization of inherited RNA-protein complexes, and discover the molecular purpose of inheriting these protein complexes. This interest in inherited RNA binding proteins currently extends to investigating RNA-protein complexes that form multi-component RNA-protein granules, or biomolecular condensates. Much of my previous work centered on characterizing the structural organization of P granules, an inherited RNA-protein granule necessary for C. elegans nematode germ cell development. The next five years will focus on understanding the molecular mechanisms how RNA-protein complexes are inherited across cell generations and from parent to progeny. Which RNA binding proteins are inherited, and which cells inherit these proteins? What protein attributes are required for inheritance? Our ability to investigate these questions is currently limited by available methods to track protein components in multicellular organisms. My lab seeks to label and follow maternal proteins in the authentic germline tissue of C. elegans, a proven model organism to study basic questions in animal development. Established single gene editing methods, robust imaging capabilities, and short generational time make C. elegans an ideal multicellular animal to identify the functions of maternal proteins inherited across generations. Modified enzyme tags now allow us to pulse label proteins with covalently bound ligands and chase these labeled proteins over time. This in vivo pulse-chase method has been used to follow chromatin remodeling in cell culture and protein stability in mouse tumors. Our preliminary results demonstrate that we are able to use in vivo pulse chase to track histone protein stability in worm germline tissue under different nutrient conditions. The current goal is to use in vivo pulse chase in C. elegans to visualize the stability of maternal germline RNA binding proteins through germ cell development and track these proteins as they are inherited from mother to progeny. First, we will pulse-label a P granule assembly protein and test how granule formation and protein quantity affect its inheritance. Second, we will pulse-chase maternal germline Argonautes, a family of RNA regulatory enzymes, to identify which Argonautes are inherited by progeny, what tissues inherit them, and what protein attributes are necessary for tissue-specific inheritance. Collectively, this work will redefi...